PUBLICATION
Novel Vector Systems Optimized for Injecting in vitro-synthesized mRNA into Zebrafish Embryos
- Authors
- Ro, H., Soun, K., Kim, E.J., and Rhee, M.
- ID
- ZDB-PUB-040609-13
- Date
- 2004
- Source
- Molecules and cells 17(2): 373-376 (Journal)
- Registered Authors
- Ro, Hyunju
- Keywords
- none
- MeSH Terms
-
- Animals
- Genes, Reporter
- Genetic Vectors*
- In Situ Hybridization
- Microinjections
- RNA, Messenger/metabolism*
- Transfection/methods*
- Zebrafish/anatomy & histology
- Zebrafish/embryology*
- Zebrafish/genetics
- PubMed
- 15179057
Citation
Ro, H., Soun, K., Kim, E.J., and Rhee, M. (2004) Novel Vector Systems Optimized for Injecting in vitro-synthesized mRNA into Zebrafish Embryos. Molecules and cells. 17(2):373-376.
Abstract
Microinjection of nucleic acids or proteins is a useful way of studying embryonic development. In particular, injection of in vitro-transcribed capped RNA is commonly employed to achieve ectopic or increased expression of genes. Two vector systems, pCS2+ and pT7Ts, have been used for this purpose in zebrafish. However, they were initially optimized for Xenopus embryos not for zebrafish. Here we describe a vector, pcGlobin2, optimized for zebrafish, and its derivative, pcGlobin2-GST. This new vector system offers several advantages. First, pcGlobin 2 contains three critical elements [5 cent and 3 cent zebrafish b-globin UTRs, and a poly(A) tail] for generating stable mRNAs and greatly improving translation efficiency. Second, subcloning and preparation of template DNA is easier because of the larger number of restriction sites. Third, protein-binding assays can be performed directly on the injected embryos with pcGlobin2-GST. Lastly, this vector system can be transfected into animal cells without additional subcloning.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping